Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs exhibit a dual role in the progression of osteoarthritis. (A) Overview of animal experiments, mice were randomly sorted into 3 groups (Sham, Control, MDSC, n = 6). The MDSC group received intra-articular injections of MDSCs twice a week, and the animals were sacrificed one and four weeks after surgery. (B) Micro-CT images of knee joint after injected MDSCs. (C) The relative joint space. (D–E) Safranin O-fast green, H&E, (F) The Mankin scores of knee joints. (G) COL2A1, and MMP-13 staining of knee joints. (H) Quantitative analysis of collagen II-positive cells and MMP-13 expression. (I) Volcano plot of differentially expressed genes (DEGs) between OA and Sham. (J) Heat map of differentially expressed genes between OA and sham. (K) Volcano plot of differentially expressed genes (DEGs) between OA and MDSC. (L) The gene set enrichment analysis (GSEA) of MDSC. (M) The kyoto encyclopedia of genes and genomes (KEGG) analysis of MDSC. (N) The expression levels of various IL-17 receptors between OA and MDSC. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Control, Micro-CT, Injection, Staining, Expressing

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs exacerbate arthritis by promoting Th17 differentiation, but depleting MDSCs can alleviate the progression of osteoarthritis. (A) One week after the injection of MDSC, compared to the control group, the intra-articular levels of TNF-α, IFN-γ, IL-1β, and IL-6. (B) Flow cytometry showed the expression of IL-17A in the joints one week after the DMM surgery. One week after the injection of MDSC, compared to the control group, the intra-articular concentrations of IL-17 (C) in serum (D) in joint. (E) Immunofluorescence staining of CD4 and IL-17 in the joints and synovial membrane of mice one week after DMM surgery. (F) Immunofluorescence of COL2A1 and MMP-13 in ATDC5 cells co-cultured with Th17 cells for four days. (G) Quantitative analysis of immunofluorescence for COL2A1 and MMP-13. (H) Flow cytometry analysis of IL-17 expression and IL-17 levels in the culture supernatant after co-culturing MDSCs with CD4 + T cells for four days, compared to the control group. Mice were randomly sorted into 3 groups (Sham, Control, Anti-Gr-1, n = 6). The Anti-Gr-1 group received i.p. injected anti-Gr-1 antibodies (100 μg) twice a week, and animals were sacrificed four weeks after surgery. (I) Micro-CT images of knee joint after injection of anti-Gr-1 at 4 weeks. (J) Quantitative results of bone mineral density after injection of anti-Gr-1. (K) Safranin O-fast green, H&E staining of knee joints after injection of anti-Gr-1 at 4 weeks. (L) Total OARSI score in different groups after injection of anti-Gr-1. (T: tibial, F: femoral) (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Injection, Control, Flow Cytometry, Expressing, Immunofluorescence, Staining, Membrane, Cell Culture, Micro-CT

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs accelerate cartilage regeneration by regulating macrophage M2 polarization. (A) Flow cytometry analysis of CD206 expression in joint. (B) Flow cytometry analysis of CD86 expression in joint. (C) One week after the injection of MDSCs, immunofluorescence of F4/80, CD86, and CD206 in the synovial membrane (n = 3). (D) Flow cytometriy analysis of CD206 expression after co-culturing MDSCs with RAW264.7 cells. (E) Cytokines levels of M2 macrophage-in the supernatant of the culture medium after co-culturing MDSCs with RAW264.7 cells. (F) Immunofluorescence staining of COL2A1 and MMP-13 after co-culturing M2 macrophages with ATDC5 cells for four days. (G) Quantitative analysis of immunofluorescence for COL2A1 and MMP-13. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Flow Cytometry, Expressing, Injection, Immunofluorescence, Membrane, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β microspheres accelerate cartilage repair by inhibiting Th17 differentiation. (A) MDSCs regulate the differentiation of CD4 + T cells and macrophages. (B) Flow cytometric analysis of IL-17 expression after co-culturing microspheres with CD4 + T cells for four days (n = 3). (C) Quantitative analysis of flow cytometry results and the concentration of IL-17 in the culture supernatant. After co-culturing MDSCs with CD4 T cells, collect the culture medium to treat ATDC5 cells and perform immunofluorescence staining of cartilage-related markers after co-culturing for four days with different groups of microspheres. (D) Immunofluorescence staining of COL2A1 (n = 6). (E) Immunofluorescence staining of MMP-13 (n = 6). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Expressing, Flow Cytometry, Concentration Assay, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β microspheres accelerate cartilage repair by promoting M2 polarization of macrophages. (A) Flow cytometric analysis of CD206 expression after co-culturing microspheres with Raw264.7 cells. (B) After pre-treating ATDC5 cells with TNF-α for one day, co-culture them with M2 polarized macrophages for four days, followed by immunofluorescence staining of MMP-13. (C) Quantitative analysis of MMP-13 immunofluorescence (n = 6). (D) After pre-treating ATDC5 cells with TNF-α for one day, co-culture them with M2 polarized macrophages for four days, followed by immunofluorescence staining of COL2A1. (E) Quantitative analysis of COL2A1 immunofluorescence (n = 6). (F) The effect of HAMA-MSN@TGF-β1/Anti-IL-1β microspheres on the expression of genes related to cartilage metabolism (n = 3). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Expressing, Co-Culture Assay, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β alleviates cartilage loss. Mice were randomly sorted into 5 groups (Sham, Control, HAMA-MSN@TGF-β1, HAMA-MSN@Anti-IL-1β, HAMA-MSN@TGF-β1/Anti-IL-1β). (A) X-ray and Micro-CT images of the mice joints at week eight. (B) Quantitative analysis of joint space and subchondral bone density (n = 3). (C) Representative microscopic photographs of Safranine O/Fast Green and H&E staining. the areas in the black squares are magnified and shown separately. (red: cartilage proteoglycan; green: collagen of subchondral bone). (D) Total OARSI score in different groups. (E) Representative microscopic photographs of COL2A1 and MMP-13 immunohistochemical staining. (F, G) Quantitative analysis of MMP-13 and COL2A1 expression (n = 3). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Control, Micro-CT, Staining, Immunohistochemical staining, Expressing